CN114175978A - Extraction method of celery root secretion and application of celery root secretion in inducing tomatoes to generate anti-bemisia tabaci defense reaction - Google Patents
Extraction method of celery root secretion and application of celery root secretion in inducing tomatoes to generate anti-bemisia tabaci defense reaction Download PDFInfo
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Abstract
The invention relates to an extraction method of celery root exudates and application of the celery root exudates in inducing tomatoes to generate a bemisia tabaci defense reaction. The invention is expected to realize green prevention and control of bemisia tabaci, and has important theoretical and practical significance for policy implementation of 'weight reduction and drug reduction' and green comprehensive prevention and control of plant diseases and insect pests and food safety construction.
Description
Technical Field
The invention relates to a technology for inducing tomatoes to generate a bemisia tabaci defense reaction by celery root exudates, in particular to an extraction method of the celery root exudates and an application of the celery root exudates in inducing the tomatoes to generate the bemisia tabaci defense reaction, and belongs to the technical field of green prevention and control of pests.
Background
Celery (Apium graveolens L.) Is a two-year-old herbaceous plant of Apium of Umbelliferae, is an important green vegetable in China, is praised as a good vegetable medicine, and has easy cultivation and developed root system. The widely planted species in China include Tianjin solid celery, Chinese herb celery and the like. Bemisia tabaci (Bemisia tabaci) Is an external invasive pest with serious harmfulness, has a wide host range of about 600 species and has a continuously increasing trend. Bemisia tabaci harms hosts by directly sucking plant sap, spreading (more than 100) viruses and secreting honeydew to cause sooty mould.
Tomato (A)Lycopersicon esculentum Mill ) The tomato cultivation area and the total yield of China all live at the top of the world, but the tomato cultivation area and the total yield of China are used as hosts with strong preference for bemisia tabaci, so that great loss is caused to tomato production due to harm of the tomato cultivation area and the tomato cultivation area, and the tomato cultivation area and the total yield become main obstacles for limiting the development of tomato industry in China. At present, the main method for preventing and controlling tomato bemisia tabaci in production still mainly applies chemical pesticide. The long-term use of chemical pesticides brings about various disadvantages: environmental pollution, increasingly enhanced drug resistance of bemisia tabaci, harm to human bodies caused by food safety due to pesticide residues, killing of beneficial organisms (particularly natural enemy organisms) and further reduction of biological diversity in the nature, and the like.
Disclosure of Invention
The invention aims to solve the problems of harm of bemisia tabaci (especially Q-type bemisia tabaci) and defects caused by a method for mainly preventing and treating chemical pesticides in the existing tomato planting industry, provides a method for treating tomatoes by using celery root exudates to generate induced insect resistance, realizes the generation of the insect resistance of the tomatoes to prevent and control the harm of the bemisia tabaci, avoids adverse effects caused by environment, food, insect pest prevention and treatment and the like caused by the use of chemical agents, and particularly relates to an extraction method of the celery root exudates and application of the method for inducing the tomatoes to generate anti-bemisia tabaci defensive reaction.
The purpose of the invention is realized as follows: the extraction method of celery root secretion is characterized by comprising the following steps:
1) cultivating celery seedlings: celery is cultivated in a plug tray, 50 celery seeds are sown in each tray, the celery seeds are cultivated in an artificial climate chamber, artificial illumination is adopted for 12 hours, and the environmental conditions of day and night temperature difference of 20 ℃/15 ℃ and relative humidity of 60-80% are adopted;
2) and pot culture: transplanting the celery seedlings into a nutrition pot till the early growth stage of the celery, wherein the seedlings are transplanted into the nutrition pot one by one, and the celery seedlings are transplanted into the nutrition pot under the conditions that the temperature difference between day and night is 20 ℃/15 ℃, and the illumination time L is D = 14: 10. continuously culturing under the environmental condition with the relative humidity of 60-80 percent;
3) transitional culture before water culture of celery: celery grows to take out its from nutrition bowl care when middle period, avoids hindering the root, gets the robust unanimous celery of growth vigor, gets rid of adnexed soil on the root, carries out the transitional culture in inserting its root system part in the light-resistant water planting box that is equipped with the pure water, puts into the incubator after 3 days and cultivates, and the cultivation environmental condition is: day and night temperature difference of 20 ℃/15 ℃, and illumination time L, D = 14: 10. relative humidity is 60% -80%, and oxygen is introduced at the same time, wherein the oxygen introduction amount is 2 multiplied by 2.5L/min;
4) extracting celery root exudate to obtain a celery root exudate solution;
celery root exudate generation mode 1: after the transitional cultivation, celery root system can grow more prosperous, with the light-resistant water planting box of celery root system part insertion pure water, put into the artificial climate case and carry out the water planting and cultivate, the cultivation ambient condition is: day and night temperature difference of 20 ℃/15 ℃, and illumination time L, D = 14: 10. relative humidity is 60% -80%; after the celery is hydroponically cultured for 7 days, collecting the solution in the hydroponic box, and storing the solution at 4 ℃ for later use;
celery root exudate generation mode 2: extracting by using a soil culture elution mode.
In the step 2), when celery is cultured to the middle growth stage, removing other impurities on the surface of celery root system soil, wherein the other impurities comprise plant debris, larger hard soil particles and vermiculite, and collecting fresh roots and surrounding soil of the celery; sieving with 20 mesh sieve, and keeping fine soil particles; adding deionized water into fresh rhizosphere soil, shaking and leaching, collecting eluent cultured by celery root system soil, and storing at 4 ℃ for later use.
In the step 1), the celery variety is Chinese cress;
in the step 1), the components in the plug are grass carbon and vermiculite, and the ratio of the grass carbon to the vermiculite is 3: 1;
in the step 2), the nutrition pot comprises grass peat and vermiculite, wherein the ratio of the grass peat to the vermiculite is 3: 1.
in the step 2), the celery taken out from the nutrition pot is 20cm-30cm in height.
The celery root exudate extracted by the extraction method of the celery root exudate is used for inducing and improving the defense reaction of tomatoes against bemisia tabaci.
The method for verifying the effect of inducing tomato to resist bemisia tabaci by using the celery root exudates extracted by the method for extracting the celery root exudates comprises the following steps:
1) the method comprises the following steps of (1) accelerating germination of tomato seeds to be tested, sowing the tomato seeds into a plug tray, and selecting robust tomatoes with similar growth vigor to transplant into nutrition pots when seedlings grow to 2-3 true leaves;
2) and culturing in an artificial climate box under the climate conditions of the temperature of 23 +/-1 ℃, the relative humidity of 60-80%, the illumination time L: D = 14: 10, uniformly managing water and fertilizer, and testing when the tomatoes grow to 4-6 true leaves;
3) cleaning the tomato roots, and inserting the cleaned tomato roots into the celery root exudate solution for water culture; using pure water culture as control, repeating the treatment for 5 times, and repeating for 4 tomatoes;
4) after the tomatoes are cultured for 7 days, the tobacco whitefly is inoculated on the leaves, and the selection behavior, the egg laying amount and the growth and development conditions of the tobacco whitefly on the pure water control CK and celery root secretion water culture tomatoes are observed;
the observation result shows that compared with the pure water CK treatment, the selectivity of the tomato plant by the bemisia tabaci on the tomato treated by the celery root exudates is obviously reduced, the egg laying amount is obviously reduced, and the growth and development are obviously inhibited.
The method is advanced and scientific, and the following technical scheme is adopted by the invention: extracting celery root secretion according to the extraction method of the celery root secretion, wherein the celery root secretion is used for inducing and improving tomato bemisia tabaci resistance.
The extraction method of celery root secretion comprises the following steps: step 1) cultivation of celery seedlings: celery is cultivated in a plug tray, 50 celery seeds are sown in each tray, the celery seeds are cultivated in an artificial climate chamber, artificial illumination is adopted for 12 hours, and the environmental conditions that the temperature difference between day and night is 20 ℃/15 ℃ and the relative humidity is 60-80% are adopted. Step 2) pot culture: transplanting the celery seedlings into a nutrition pot till the early growth stage of the celery, wherein the seedlings are transplanted into the nutrition pot one by one, and the celery seedlings are transplanted into the nutrition pot under the conditions that the temperature difference between day and night is 20 ℃/15 ℃, and the illumination time L is D = 14: 10. the culture is continued under the environmental condition with the relative humidity of 60 percent to 80 percent. Step 3) transitional culture before water culture of celery: carefully taking out celery from the nutrition pot when the celery grows to the middle stage (avoiding damaging roots), removing the robust and consistent celery, removing soil attached to the roots, inserting the root system part of the celery into a light-proof water culture box filled with pure water for transitional culture, putting the celery into an incubator for culture after 3 days, and introducing oxygen under the same environmental condition as the step 2) with the oxygen introduction amount of 2 multiplied by 2.5L/min. Step 4), celery root exudate: production mode 1: after the transitional culture is finished, the celery root system can grow vigorously, the celery root system part is inserted into a pure water lucifugal water culture box and is placed into an artificial climate box for water culture, and the provided growth conditions are the same as those in the step 3); after the celery is hydroponically cultured for 7 days, collecting the solution in the hydroponic box, and storing the solution at 4 ℃ for later use; celery root exudate generation mode 2: extracting by using a soil culture elution mode. When the celery in the step 2) is cultured to the middle growth stage, removing other impurities (plant debris, larger hard soil particles, vermiculite and the like) on the surface of the root system soil of the celery, and collecting fresh root and surrounding soil of the celery. Sieving with 20 mesh sieve, and keeping fine soil particles; adding deionized water into fresh rhizosphere soil, shaking and leaching, collecting eluent cultured by celery root system soil, and storing at 4 ℃ for later use.
The method for verifying the bemisia tabaci resistant effect of the celery root secretion induced tomatoes comprises the following steps: step 1) accelerating germination of tomato seeds to be tested, sowing the tomato seeds into a plug tray, selecting strong tomatoes with similar growth vigor to transplant into nutrition pots when the seedlings grow to 2-3 true leaves, and transplanting 1 tomato plant in each pot. Step 2) culturing in an artificial climate box under the climate conditions of temperature 23 +/-1 ℃, relative humidity 60% -80%, illumination time L: D = 14: and 10, uniformly managing water and fertilizer, and testing when the tomatoes grow to 4-6 true leaves. And 3) cleaning the tomato roots, and inserting the cleaned tomato roots into the celery root exudate solution for water culture. The treatment was repeated 5 times, 4 tomatoes per repeat, using pure water hydroponics as a control. And 4) inoculating bemisia tabaci on leaves after the tomatoes are cultured for 7 days, and observing the selective behavior, the oviposition amount, the development duration and the like of the bemisia tabaci on the pure water control CK and celery root secretion water culture tomatoes. The observation result shows that compared with the pure water CK treatment, the selectivity of the celery root secretion for treating the bemisia tabaci on the tomatoes is obviously reduced, the egg laying amount is obviously reduced, and the growth and development are obviously inhibited.
The invention uses celery (such as Chinese herb celery) root secretion to treat tomato plants, induces tomato to generate insect resistance, activates autoimmune system, achieves the purpose of protecting vegetables, and further avoids or is expected to greatly reduce the application of chemical pesticides and avoid the generation of the defects. The invention is expected to realize green prevention and control of bemisia tabaci, and has important theoretical and practical significance for policy implementation of 'weight reduction and drug reduction' and green comprehensive prevention and control of plant diseases and insect pests and food safety construction.
Drawings
FIG. 1 shows the egg laying amount of bemisia tabaci on tomato leaves treated with celery root exudates in the present invention; the data in FIG. 1 are mean. + -. standard error. Different letters indicate significant differences between treatments (p < 0.05).
FIG. 2 shows the selectivity of Bemisia tabaci on tomato plants treated with celery root exudate in accordance with the present invention; note: the data in FIG. 2 are mean. + -. standard error. Different letters indicate significant differences between treatments (p < 0.05).
Detailed Description
The invention is further described with reference to the accompanying drawings and the description thereof.
A celery root secretion for inducing and improving the defense reaction of tomato against bemisia tabaci is disclosed. The extraction method of celery root secretion comprises the following steps:
1) cultivating celery seedlings: celery is cultivated in a plug tray, 50 celery seeds are sown in each tray, the celery seeds are cultivated in an artificial climate chamber, artificial illumination is adopted for 12 hours, and the environmental conditions of day and night temperature difference of 20 ℃/15 ℃ and relative humidity of 60-80% are adopted;
2) pot culture: transplanting the celery seedlings into a nutrition pot till the early growth stage of the celery, wherein the seedlings are transplanted into the nutrition pot one by one, and the celery seedlings are transplanted into the nutrition pot under the conditions that the temperature difference between day and night is 20 ℃/15 ℃, and the illumination time L is D = 14: 10. continuously culturing under the environmental condition with the relative humidity of 60-80 percent;
3) transitional culture before water culture of celery: when celery grows to the middle period, carefully taking out the celery from the nutrition pot (avoiding injuring the roots), removing the robust and consistent celery, removing the soil attached to the roots, inserting the root system part of the celery into a light-resistant water culture box filled with pure water for transitional culture, and putting the celery into an incubator for culture after 3 days, wherein the environmental conditions are the same as the step 2). Simultaneously introducing oxygen with the oxygen flow rate of 2 multiplied by 2.5L/min;
4) celery root secretion: production mode 1: after the transitional culture is finished, the celery root system can grow vigorously, the celery root system part is inserted into a light-tight water culture box of pure water and is put into an artificial climate box for water culture, and the provided growth conditions are the same as the step 3); after the celery is hydroponically cultured for 7 days, collecting the solution in the hydroponic box, and storing the solution at 4 ℃ for later use; celery root exudate generation mode 2: extracting by using a soil culture elution mode.
When the celery in the step 2) is cultured to the middle growth stage, removing other impurities (plant debris, larger hard soil particles, vermiculite and the like) on the surface of the root system soil of the celery, and collecting fresh root and surrounding soil of the celery. Sieving with 20 mesh sieve, and keeping fine soil particles; adding deionized water into fresh rhizosphere soil, shaking and leaching, collecting eluent cultured by celery root system soil, and storing at 4 ℃ for later use.
In the step 1), the celery variety is Chinese medicine celery, and the plug tray in the step 1) and the nutrition pot in the step 2) adopt the following components and proportions of grass carbon: vermiculite = 3: 1. in the step 2), the celery taken out from the nutrition pot is 20cm-30cm in height.
The method for verifying the bemisia tabaci resistant effect of the celery root secretion induced tomatoes comprises the following steps:
1) accelerating germination of tomato seeds to be tested, sowing the tomato seeds in a plug tray, selecting robust tomatoes with similar growth vigor to transplant into nutrition pots when the seedlings grow to 2-3 true leaves, and transplanting 1 tomato in each pot;
2) culturing in an artificial climate box under the climate conditions of temperature 23 +/-1 ℃, relative humidity 60-80%, illumination time L: D = 14: 10, uniformly managing water and fertilizer, and testing when the tomatoes grow to 4-6 true leaves;
3) cleaning tomato roots, and inserting the cleaned tomato roots into the celery root exudate solution for water culture. Using pure water culture as control, repeating the treatment for 5 times, and repeating for 4 tomatoes;
4) after the tomatoes are cultured for 7 days, the tobacco whitefly is inoculated on the leaves, and the selective behavior, the egg laying amount and the growth and development conditions of the tobacco whitefly on the pure water control CK and celery root secretion water culture tomatoes are observed. The observation result shows that compared with the pure water CK treatment, the selectivity of the celery root secretion for treating the bemisia tabaci on the tomatoes is obviously reduced, the egg laying amount is obviously reduced, and the growth and development are obviously inhibited.
Example 1, oviposition of bemisia tabaci on celery root exudate treated tomato leaves was performed as follows:
placing celery in an environment with the temperature difference of 20/15 ℃ day and night for soil culture and water culture respectively. Extracting the celery hydroponic solution: selecting 12 seedlings with good growth vigor, cleaning with deionized water to remove root soil, placing in pure water for 3 days for transition, and culturing in pure water to ensure normal growth of the plants. After culturing for 1 week, collecting root secretion sample, filtering quickly to collect the solution in the water culture box, and storing in a refrigerator at 4 deg.C. Extracting the celery soil culture eluent: the ready-to-sterilize shovel performs the collection of the soil, removing other impurities (larger hard soil particles, plant debris, etc.) from the soil surface prior to collecting the soil. Sieving with 20 mesh sieve to remove plant root system, animal debris and other residues. Adding deionized water into fresh rhizosphere soil, shaking for leaching, rapidly filtering to collect solution in the water culture box, and storing in a refrigerator at 4 deg.C. Tomato treatment: tomato roots were washed and inserted into celery root exudate of different collection modes for culture for 7 days, pure water culture was used as control CK treatment, and each treatment was repeated 5 times. In addition, tomato 'Pindou 53' is planted in a plug tray of an artificial climate chamber, when the tomato grows to 3-4 true leaves, tomato seedlings which grow closely and are healthy and free of diseases and pests are taken and inserted into a light-resistant water culture box filled with celery root exudates, and the control group is treated by pure water. The tomato plants of the experimental group and the tomato plants of the control group are placed into a circle with the diameter of 1 meter and placed in a cage made of gauze. The bemisia tabaci is released at night and released in the center of the net cage above the plant canopy. The sampling bottle assembly is filled with bemisia tabaci and hung in a transparent plastic glass in the center of the cage, and the distance between the bemisia tabaci and the transparent plastic glass is about 30cm above the plant canopy. The bemisia tabaci moves to the open top, flies out of the sample bottle and approaches the plant from above. The test tomatoes were taken out after 24 hours, and the oviposition of bemisia tabaci was counted under a scope of a scope. The experiment was repeated 5 times. As can be seen from figure 1, the oviposition of bemisia tabaci in the control CK group of pure water is large, and the oviposition of the celery root exudates on the leaves of the tomato group treated by celery is obviously reduced. The celery root secretion treatment effectively inhibits the egg laying of bemisia tabaci on the tomatoes, so that the aim of inducing and improving the resistance of the tomatoes to the bemisia tabaci is fulfilled.
Example 2, the selection behavior of bemisia tabaci on celery root exudate treatment tomato plants was performed as follows:
placing celery in an environment with the temperature difference of 20/15 ℃ day and night for soil culture and water culture respectively. Extracting the celery hydroponic solution: selecting 12 seedlings with good growth vigor, cleaning with deionized water to remove root soil, placing in pure water for 3 days for transition, and culturing in pure water to ensure normal growth of the plants. After culturing for 1 week, collecting root exudates, and storing in a refrigerator at 4 deg.C. Extracting the celery soil culture eluent: a shovel prepared for sterilization is used for collecting soil, and plant debris, larger and harder soil particles and the like on the surface of the soil are removed before the soil is collected). Sieving with 20 mesh sieve to remove obvious plant root system, animal debris and other large impurities. Adding deionized water into fresh rhizosphere soil, shaking for leaching, rapidly filtering, collecting root secretion solution, and storing in a refrigerator at 4 deg.C. Treating the tomato with celery root secretion: in an artificial climate room, planting tomato 'Pindu 53' in a plug tray, when the tomato grows to 3-4 true leaves, taking tomato seedlings which grow closely and are healthy and free of diseases and pests, inserting the tomato seedlings into a light-resistant water culture box filled with 1000mL of celery root exudates, and culturing, wherein pure water is used as a control CK. All tomato plants in the treated group and the control group were placed in a round shape with a diameter of 1 m and placed in a cage made of gauze. The other experimental procedures were the same as in "example 1". And taking out after 24 hours, and immediately investigating the number of bemisia tabaci on the tomato leaves. The experiment was repeated 5 times. As can be seen from FIG. 2, the number of bemisia tabaci on tomato plants in the pure water CK group was greater, while the number of leaf plants in the celery root exudate treated tomato group was significantly reduced. The treatment of the celery root secretion effectively inhibits the number of bemisia tabaci on the tomatoes, so that the aim of inducing and improving the resistance of the tomatoes to the bemisia tabaci is fulfilled.
Claims (6)
1. The extraction method of celery root secretion is characterized by comprising the following steps:
1) cultivating celery seedlings: celery is cultivated in a plug tray, 50 celery seeds are sown in each tray, the celery seeds are cultivated in an artificial climate chamber, artificial illumination is adopted for 12 hours, and the environmental conditions of day and night temperature difference of 20 ℃/15 ℃ and relative humidity of 60-80% are adopted;
2) and pot culture: transplanting the celery seedlings into a nutrition pot till the early growth stage of the celery, wherein the seedlings are transplanted into the nutrition pot one by one, and the celery seedlings are transplanted into the nutrition pot under the conditions that the temperature difference between day and night is 20 ℃/15 ℃, and the illumination time L is D = 14: 10. continuously culturing under the environmental condition with the relative humidity of 60-80 percent;
3) transitional culture before water culture of celery: celery grows to take out its from nutrition bowl care when middle period, avoids hindering the root, gets the robust unanimous celery of growth vigor, gets rid of adnexed soil on the root, carries out the transitional culture in inserting its root system part in the light-resistant water planting box that is equipped with the pure water, puts into the incubator after 3 days and cultivates, and the cultivation environmental condition is: day and night temperature difference of 20 ℃/15 ℃, and illumination time L, D = 14: 10. relative humidity is 60% -80%, and oxygen is introduced at the same time, wherein the oxygen introduction amount is 2 multiplied by 2.5L/min;
4) extracting celery root exudate to obtain a celery root exudate solution;
celery root exudate generation mode 1: after the transitional cultivation, celery root system can grow more prosperous, with the light-resistant water planting box of celery root system part insertion pure water, put into the artificial climate case and carry out the water planting and cultivate, the cultivation ambient condition is: day and night temperature difference of 20 ℃/15 ℃, and illumination time L, D = 14: 10. relative humidity is 60% -80%; after the celery is hydroponically cultured for 7 days, collecting the solution in the hydroponic box, and storing the solution at 4 ℃ for later use;
celery root exudate generation mode 2: extracting by using a soil culture elution mode.
2. The method for extracting celery root exudates according to claim 1, wherein in the step 2), when celery is cultured to the middle growth stage, other impurities on the surface of celery root soil are removed, wherein the other impurities comprise plant debris, large hard soil particles and vermiculite, and fresh roots and surrounding soil of the celery root soil are collected; sieving with 20 mesh sieve, and keeping fine soil particles; adding deionized water into fresh rhizosphere soil, shaking and leaching, collecting eluent cultured by celery root system soil, and storing at 4 ℃ for later use.
3. The method for extracting celery root exudates according to claim 1, wherein in the step 1), the celery is Chinese parsley;
in the step 1), the components in the plug are grass carbon and vermiculite, and the ratio of the grass carbon to the vermiculite is 3: 1;
in the step 2), the nutrition pot comprises grass peat and vermiculite, wherein the ratio of the grass peat to the vermiculite is 3: 1.
4. the method for extracting celery root exudates according to claim 1, wherein in the step 2), the celery taken out of the nutrition pot is 20cm-30cm high.
5. The celery root exudate extracted by the method for extracting the celery root exudate as claimed in any one of claims 1 to 4 is used for inducing and improving the defense response of tomatoes against bemisia tabaci.
6. The method for verifying the effect of inducing tomato to resist bemisia tabaci by the celery root exudates extracted by the method for extracting the celery root exudates as claimed in any one of claims 1 to 4 is characterized by comprising the following steps of:
1) the method comprises the following steps of (1) accelerating germination of tomato seeds to be tested, sowing the tomato seeds into a plug tray, and selecting robust tomatoes with similar growth vigor to transplant into nutrition pots when seedlings grow to 2-3 true leaves;
2) and culturing in an artificial climate box under the climatic conditions of the temperature of 23 +/-1 ℃, the relative humidity of 60-80 percent and the illumination time L, D = 14: 10, uniformly managing water and fertilizer, and testing when the tomatoes grow to 4-6 true leaves;
3) cleaning the tomato roots, and inserting the cleaned tomato roots into the celery root exudate solution for water culture; using pure water culture as control, repeating the treatment for 5 times, and repeating for 4 tomatoes;
4) after the tomatoes are cultured for 7 days, the leaf blades are inoculated with the bemisia tabaci, and the selective behavior, the egg laying amount and the development period of the bemisia tabaci on the pure water control CK and the celery root secretion water culture tomatoes are observed;
the observation result shows that compared with pure water CK treatment, the selectivity of the bemisia tabaci on the tomato treated by the celery root exudates is obviously reduced, the egg laying amount is obviously reduced, and the growth and development of the tomato at each age are obviously inhibited.
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